Secretion of the alpha-galactosidase from Cyamopsis tetragonoloba (guar) by Bacillus subtilis
| Autor: | G. H. M. Termorshuizen, C. T. Verrips, N. Overbeeke, D. R. Underwood, M. L. F. Giuseppin |
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| Rok vydání: | 1990 |
| Předmět: |
Glycosylation
Bacillus amyloliquefaciens Cyamopsis Blotting Western Molecular Sequence Data Guar Bacillus subtilis Applied Microbiology and Biotechnology chemistry.chemical_compound Amino Acid Sequence Gel electrophoresis Plants Medicinal Guar gum Ecology biology fungi Fabaceae biology.organism_classification Enzyme assay Galactosidases Biochemistry chemistry alpha-Galactosidase Fermentation biology.protein Transformation Bacterial Research Article Plasmids Food Science Biotechnology |
| Zdroj: | Applied and Environmental Microbiology. 56:1429-1434 |
| ISSN: | 1098-5336 0099-2240 |
| DOI: | 10.1128/aem.56.5.1429-1434.1990 |
| Popis: | A fusion of DNA sequences encoding the SPO2 promoter, the alpha-amylase signal sequence from Bacillus amyloliquefaciens, and the mature part of the alpha-galactosidase from Cyamopsis tetragonoloba (guar) was constructed on a Bacillus subtilis multicopy vector. Bacillus cells of the protease-deficient strain DB104 harboring this vector produced and secreted the plant enzyme alpha-galactosidase up to levels of 1,700 U/liter. A growth medium suppressing the residual proteolytic activity of strain DB104 was used to reach these levels in a fermentor. Purification of the secreted product followed by NH2-terminal amino acid sequencing showed that the alpha-amylase signal sequence had been processed correctly. The molecular mass of the product estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was slightly lower than that of the plant purified enzyme, which is most likely due to glycosylation of the latter. The alpha-galactosidase product was active both on the artificial substrate para-nitrophenyl-alpha-D-galactopyranoside and on the galactomannan substrate, guar gum. The activity of this Bacillus sp.-produced enzyme was similar to that of the glycosylated enzyme purified from guar seeds, indicating that glycosylation has no essential function for enzyme activity. |
| Databáze: | OpenAIRE |
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